Background: EGFR exon 20 insertions account for 1% to 10% of EGFR mutations in nonsmall-cell lung cancer (NSCLC) and are known to confer resistance to traditional tyrosine kinase inhibitors. However, the prognostic significance of these mutations in early-stage resected NSCLC remains unclear. This study assesses outcomes in patients with resected NSCLC harboring EGFR exon 20 insertions, comparing them to patients with common EGFR mutations and those with wild-type EGFR. Methods: We retrospectively reviewed 3,235 patients who underwent resection for NSCLC at Tri-Service Hospital between 2008 and 2021. After excluding cases lacking EGFR testing, incomplete data, or advanced-stage disease, 44 patients with exon 20 insertions were matched to 602 patients with common EGFR mutations and 729 with wild-type EGFR. Clinical characteristics, disease-free survival (DFS), and overall survival (OS) were analyzed using the Kaplan-Meier method, with statistical comparisons performed using the log-rank test. Cox proportional hazards models were used to identify independent prognostic factors. Results: Patients with exon 20 insertions were younger and more frequently presented with stage IA disease. The 5-year DFS was 79% in the exon 20 insertion group, compared to 81% in the common mutation group and 83.9% in the wild-type group. The 5-year OS was 78.5% for exon 20, 91.9% for common mutations, and 91% for wild-type. While no significant differences in DFS or OS were observed between groups, the exon 20 insertion group had a higher incidence of secondary cancers. Multivariable analysis indicated that exon 20 insertion was independently associated with worse OS, but not with DFS. Conclusion EGFR exon 20 insertions do not significantly shorten DFS, but are associated with inferior OS in early-stage resected NSCLC. Given the limited treatment options, the role of adjuvant therapy warrants further investigation.

The pandemic of avian influenza viruses (AIVs) in Asia has caused enormous economic loss in poultry industry and human health threat, especially clade 2.3.4.4 H5 and H7 subtypes in recent years. The endemic chicken H6 virus in Taiwan has also brought about human and dog infections. Since wild waterfowls is the major AIV reservoir, it is important to monitor the diversified subtypes in wildfowl flocks in early stage to prevent viral reassortment and transmission. To develop a more efficient and sensitive approach is a key issue in epidemic control. In this study, we integrate multiplex reverse transcription recombinase polymerase amplification (RT-RPA) and capillary electrophoresis (CE) for high-throughput detection and differentiation of AIVs in wild waterfowls in Taiwan. Four viral genes were detected simultaneously, including nucleoprotein (NP) gene of all AIVs, hemagglutinin (HA) gene of clade 2.3.4.4 H5, H6 and H7 subtypes. The detection limit of the developed detection system could achieve as low as one copy number for each of the four viral gene targets. Sixty wild waterfowl field samples were tested and all of the four gene signals were unambiguously identified within 6 h, including the initial sample processing and the final CE data analysis.The results indicated that multiplex RT-RPA combined with CE was an excellent alternative for instant simultaneous AIV detection and subtype differentiation. The high efficiency and sensitivity of the proposed method could greatly assist in wild bird monitoring and epidemic control of poultry.